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目的探讨不同信号肽对重组B群脑膜炎奈瑟菌H因子结合蛋白(factor H-binding protein,f HBP)表达的影响。方法运用PCR方法将B群脑膜炎奈瑟菌f HBP原始信号肽分别替换为大肠埃希菌主要外膜脂蛋白(major outer membrane lipoprotein,MLP)、脂蛋白-28(lipoprotein-28,LP)和流感嗜血杆菌(Haemophilus influenzae)P4外膜蛋白信号肽,并将其分别连接至p ET-30a表达载体,转化入大肠埃希菌感受态细胞,IPTG诱导表达,SDS-PAGE鉴定蛋白表达水平。结果替换了P4、LP和MLP信号肽的重组B群脑膜炎奈瑟菌f HBP表达量较带原始信号肽的B群脑膜炎奈瑟菌f HBP相比,分别提高了2.68、1.94和2.91倍。结论替换MLP信号肽对重组B群脑膜炎奈瑟菌f HBP蛋白表达的影响最大,为提高B群脑膜炎奈瑟菌f HBP表达量的研究工作提供了新思路。
Objective To investigate the effect of different signal peptides on the expression of recombinant factor B-binding protein (f HBP) of Neisseria meningitides. Methods The primary signal peptide of Neisseria meningitidis f HBP group B was replaced by major outer membrane lipoprotein (MLP), lipoprotein-28 (LP) and Haemophilus influenzae P4 outer membrane protein signal peptide, respectively, and ligated into p ET-30a expression vector, transformed into Escherichia coli competent cells, induced by IPTG, and the protein expression level was identified by SDS-PAGE. Results The recombinant group B N. meningitidis f HBP expression with P4, LP and MLP signal peptide substitutions was increased by 2.68, 1.94 and 2.91 times, respectively, as compared to group B N. meningitidis f HBP with the original signal peptide . Conclusion The substitution of MLP signal peptide has the greatest impact on the expression of f HBP protein of Neisseria meningitidis group B, which provides a new idea for the study of f HBP expression in N. meningitidis group B.